• Title/Summary/Keyword: translational research

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miR-153 Silencing Induces Apoptosis in the MDA-MB-231 Breast Cancer Cell Line

  • Anaya-Ruiz, Maricruz;Cebada, Jorge;Delgado-Lopez, Guadalupe;Sanchez-Vazquez, Maria Luisa;Perez-Santos, Jose Luis Martin
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.5
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    • pp.2983-2986
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    • 2013
  • MicroRNAs (miRNAs) are small, non-coding RNAs (18-25 nucleotides) that post-transcriptionally modulate gene expression by negatively regulating the stability or translational efficiency of their target mRNAs. In this context, the present study aimed to evaluate the in vitro effects of miR-153 inhibition in the breast carcinoma cell line MDA-MB-231. Forty-eight hours after MDA-MB-231 cells were transfected with the miR-153 inhibitor, an MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay was utilized to determine the effects of miR-153 on cell viability. Flow cytometry analysis and assessment of caspase 3/7 activity were adopted to determine whether miR-153 affects the proliferation rates and apoptosis levels of MDA-MB-231 cells. Our results showed that silencing of miR-153 significantly inhibited growth when compared to controls at 48 hours, reducing proliferation by 37.6%, and inducing apoptosis. Further studies are necessary to corroborate our findings and examine the potential use of this microRNA in future diagnostic and therapeutic interventions.

Molecular Genetics and Diagnostic Approach of Mucolipidosis II/III

  • Sohn, Young Bae
    • Journal of mucopolysaccharidosis and rare diseases
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    • v.2 no.1
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    • pp.13-16
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    • 2016
  • Mucolipidosis (ML) II/III are autosomal recessive diseases caused by deficiency of post-translational modification of lysosomal enzymes. The mannose-6-phosphate (M6P) residue in lysosomal enzymes synthesized by N-acetylglucosamine 1-phosphotransferase (GlcNAc-phosphotransferase) serves as recognition marker for trafficking in lysosomes. GlcNAc-phosphotransferase is encoded by GNPTAB and GNPTG. Mutations in GNPTAB cause severe ML II alpha/beta and the attenuated ML III alpha/beta. Whereas mutations in GNPTG cause the ML III gamma, the attenuated type of ML III variant. For the diagnostic approaches, increased urinary oligosaccharides excretion could be a screening test in clinically suspicious patients. To confirm the diagnosis, instead of measuring the activity of GlcNAc phosphotransferase, measuring the enzymatic activities of different lysosomal hydrolases are useful for diagnosis. The activities of several lysosomal hydrolases are decreased in fibroblasts but increased in serum of the patients. In addition, the sequence analysis of causative gene is warranted. Therefore, the confirmatory diagnosis requires a combination of clinical evaluation, biochemical and molecular genetic testing. ML II/III show complex disease manifestations with lysosomal storage as the prime cellular defect that initiates consequential organic dysfunctions. As there are no specific therapy for ML to date, understanding the molecular pathogenesis can contribute to develop new therapeutic approaches ultimately.

The Localization of Cytokeratin 19 and Vimentin in Sprague Dawley Albino Rat Skin Tissue

  • Kim, Tae Keun;Kim, Yong Joo;Min, Byoung Hoon;Kim, Soo Jin
    • Applied Microscopy
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    • v.44 no.1
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    • pp.15-20
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    • 2014
  • Cytokeratin 19 (CK19) expressed in epidermis of skin, bulge region of hair follicle, outermost layer of outer root sheath and proximal and distal to bulge. Vimentin is a fibrous protein that localized in cytoplasm of fibroblast and forms cytoskeleton to maintain shape of cell and nucleus. In this study, CK19 and vimentin in skin were confirmed with light, fluorescence and transmission electron microscope. As a result, CK19 was localized epidermis, hair follicles, outer root sheath and nucleus of Merkel's cell. However, vimentin was localized some epidermis, dermis, hypodermis and nucleus of Merkel's cell. The role of CK19 is self-renewal and homeostasis in skin. Also, hair follicle regeneration and hair growth is known to be related. It is supposed that required of structural proteins that make up cytoskeleton is increased. Thereby, expression of CK19 is increased. It is considered that vimentin localized in order to stabilize structure of cell and cytoskeleton of fibroblasts. Also, CK19 and vimentin present in nuclei of Merkel's cell, and to act as a fibrous protein that make up end of a nerve fiber present in Merkel's cell and paracrine function of Merkel's cell.

Intracellular Responses of Antibody-Producing H69K-NGD Transfectoma Subjected to Hyperosmotic Pressure

  • Bae, Sung-Won;Lee, Gyun-Min
    • Journal of Microbiology and Biotechnology
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    • v.15 no.3
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    • pp.579-586
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    • 2005
  • When subjected to hyperosmotic pressure by NaCl addition, H69K-NGD transfectoma, like KR12H-2 transfectoma, displayed decreased specific growth rate (${\mu}$) and increased specific antibody productivity ($q_{Ab}$): Elevation of medium osmolality from 280 mOsm/kg to 415 mOsm/kg decreased ${\mu}$ by $79\%$ in batch cultures of H69K-NGD transfectoma, while it increased $q_{Ab}$ by $103\%$. However, unlike KR12H-2 tranfectoma, enhanced $q_{Ab}$ of H69K-NGD transfectoma at hyperosmolalities was not due to elevated levels of Ig mRNAs. In hyperosmotic cultures of H69K-NGD transfectoma, heavy-chain mRNA per cell was not enhanced with increasing osmolality. Hyperosmotic pressure was found to preferentially enhance immunoglobulin (Ig) translation rates of H69K-NGD transfectoma. However, under hyperosmotic pressure, the translation rate of Ig polypeptides was not enhanced as much as $q_{Ab}$. This result suggests that hyperosmotic pressure also influences the post-translational process. Taken together, the results obtained show that intracellular response of transfectomas to hyperosmotic pressure, in regard to the main intracellular steps of the antibody secretory pathway, is cell-line dependent.

Dynamic Design of a Mass-Spring Type Translational Wave Energy Converter (파력발전용 병진 질량-스프링식 파력 변환장치의 동적설계)

  • Choi, Young-Hyu;Lee, Chang-Jo;Hong, Dae-Sun
    • Journal of the Korean Society of Manufacturing Technology Engineers
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    • v.21 no.1
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    • pp.182-189
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    • 2012
  • This study suggests a dynamic design process for deciding properly design parameters of a mass-spring type Wave Energy Converter (WEC) to achieve sufficient energy conversion from wave to power generator. The WEC mechanism, in this research, consists of a rigid sprung body, a platform, suspension springs and dampers. The rigid sprung body is supported on the platform via springs and dampers and vibrates translationally in the heave direction under wave excitation. At last the resulting heave motion of the sprung body is transmitted to rotating motion of the electric generator by rack and pinion, and transmission gears. For the purpose of vibration analysis, the WEC mechanism has been simply modelled as a mass-spring-damper system under harmonic base excitation. Its maximum displacement transmissibility and steady state response can be determined by using elementary vibration theory if the harmonic ocean wave data were provided. With the vibration analysis results, the suggested dynamic design process of WEC can determine all the design parameters of the WEC mechanism, such as sprung body mass, suspension spring constant, and damping coefficient that can give sufficient relative displacement transmissibility and the associated inertia moment to drive the electric generator and transmission gears.

Recombinant Expression, Isotope Labeling, and Purification of Cold shock Protein from Colwellia psychrerythraea for NMR Study

  • Moon, Chang-Hun;Jeong, Ki-Woong;Kim, Hak-Jun;Heo, Yong-Seok;Kim, Yang-Mee
    • Bulletin of the Korean Chemical Society
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    • v.30 no.11
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    • pp.2647-2650
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    • 2009
  • Cold shock proteins (Csps) are a subgroup of the cold-induced proteins on reduction of the growth temperature below the physiological temperature. They preferentially bind to single-stranded nucleic acids to translational regulation via RNA chaperoning. Csp plays important role in cold adaptations for the psychrophilic microorganism. Recently, Cold shock protein from psychrophilic bacteria, Colwellia psychrerythraea (CpCsp) has been identified. Three dimensional structures of a number of Csps from various microorganisms have been solved by NMR spectroscopy or X-ray crystallography, but structures of psychrophilic Csps were not studied yet. Therefore, cloning and purification protocols for further structural study of psychrophilic Csp have been optimized in this study. CpCsp was expressed in E. coli with pET-11a vector system and purified by ion exchange, size exclusion, and reverse phase chromatography. Expression and purification of CpCsp in M9 minimal media was carried out and $^{15}N$-labeled proteins with high purity over 90% was obtained. Further study will be carried out to investigate the tertiary structure and dynamics of CpCsp.

Evaluation of Joint Reaction Forces for a Hydraulic Excavator Subjected to a Critical Load (가혹하중이 작용하는 경우의 굴삭기 연결부의 반력계산)

  • Kim, Oe-Jo;Yu, Wan-Seok;Yun, Kyeong-Hwa;Gang, Ha-Geun
    • Transactions of the Korean Society of Mechanical Engineers A
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    • v.20 no.4
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    • pp.1154-1163
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    • 1996
  • This paper presents a three dimensional modeling and dynamic anlaysis of a hydraulic excavator. An excavator is composed of a ground, an under-frame, two idlers, two spockets, an upper-frame, a boom, an arm, a bucket two yokes, two connecting rods, two boom cylinders, an arm cylinder, and a bucket cylinder. Each cylinder is modeled with two separate bodies which are linked to each other by a translational joint. The three dimensioanl model of the excavator consists of 22 bodies and each body is assumed as rigid. This paper suggested the maximum lifting capability, a critical load and reaction forces at joints form the DADS simulation. It was presumed that the reaction forces due to a critical load are three times bigger than those due to the maximum lifting capacity.

Ginseng Saponin as an Antagonist for Gap Junctional Channels

  • Rhee, Seung-Keun
    • Journal of Ginseng Research
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    • v.30 no.2
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    • pp.64-69
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    • 2006
  • Gap junctional channels, allowing rapid intercellular communication and synchronization of coupled cell activities, play crucial roles in many signaling processes, including a variety of cell activities. Consequently, a modulation of the gap junctional intercellular communication (GJIC) should be a potential pharmacological target. In the present, the GJIC of a epithelial-derived rat mammary cells (BICR-M1Rk) was assessed in the presence of ginseng saponin, by using an established method of scrape-loading dye transfer assay. The transfer of Lucifer yellow (diameter: 1.2 nm) among the neighboring BICR-M1Rk cells, in which connexin43 (Cx43) is a major gap junction channel-forming protein, was significantly retarded at a concentration of $10{\mu}g/ml$ ginseng saponin. By using both methods of RT-PCR and Western blotting, it was demonstrated that ginseng saponin modulated neither the mRNA synthesis of Cx43 nor the translational process of Cx43. This ginseng saponin-induced modification of GJIC was a similar phenomenon observed under the $\beta$-glycyrrhetinic acid treatment, a well-known gap junction channel blocker. Taken together, it is reasonable to conclude that the ginseng saponin inhibits GJIC only by modulating the gating property of gap junction channels.

Genomic Insights into the Rice Blast Fungus through Estimation of Gene Emergence Time in Phylogenetic Context

  • Choi, Jaeyoung;Lee, Jong-Joon;Jeon, Junhyun
    • Mycobiology
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    • v.46 no.4
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    • pp.361-369
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    • 2018
  • The rice blast fungus, Magnaporthe oryzae, is an important pathogen of rice plants. It is well known that genes encoded in the genome have different evolutionary histories that are related to their functions. Phylostratigraphy is a method that correlates the evolutionary origin of genes with evolutionary transitions. Here we applied phylostratigraphy to partition total gene content of M. oryzae into distinct classes (phylostrata), which we designated PS1 to PS7, based on estimation of their emergence time. Genes in individual phylostrata did not show significant biases in their global distribution among seven chromosomes, but at the local level, clustering of genes belonging to the same phylostratum was observed. Our phylostrata-wide analysis of genes revealed that genes in the same phylostratum tend to be similar in many physical and functional characteristics such as gene length and structure, GC contents, codon adaptation index, and level of transcription, which correlates with biological functions in evolutionary context. We also found that a significant proportion of genes in the genome are orphans, for which no orthologs can be detected in the database. Among them, we narrowed down to seven orphan genes having transcriptional and translational evidences, and showed that one of them is implicated in asexual reproduction and virulence, suggesting ongoing evolution in this fungus through lineage-specific genes. Our results provide genomic basis for linking functions of pathogenicity factors and gene emergence time.

Vibration analysis thermally affected viscoelastic nanosensors subjected to linear varying loads

  • Ebrahimi, Farzad;Babaei, Ramin;Shaghaghi, Gholam Reza
    • Advances in nano research
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    • v.6 no.4
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    • pp.399-422
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    • 2018
  • Unwanted vibration is an issue in many industrial systems, especially in nano-devices. There are many ways to compensate these unwanted vibrations based on the results of the past researches. Elastic medium and smart material etc. are effective methods to restrain unnecessary vibration. In this manuscript, dynamic analysis of viscoelastic nanosensor which is made of functionally graded (FGM) nanobeams is investigated. It is assumed that, the shaft is flexible. The system is modeled based on Timoshenko beam theory and also environmental condition, external linear varying loads and thermal loading effect are considered. The equations of motion are extracted by using energy method and Hamilton principle to describe the translational and shear deformation's behavior of the system. Governing equations of motion are extracted by supplementing Eringen's nonlocal theory. Finally vibration behavior of system especially the frequency of system is developed by implementation Semi-analytical differential transformed method (DTM). The results are validated in the researches that have been done in the past and shows good agreement with them.